A comparison of different pre-lysis methods and extraction kits for recovery of Streptococcus agalacticae (Lancefield group B Streptococcus) DNA from whole blood

Sub-optimal recovery of bacterial DNA from whole blood samples can limit the sensitivity of molecular assays to detect pathogenic bacteria. We compared 3 different pre-lysis protocols (none, mechanical pre-lysis and achromopeptidase pre-lysis) and 5 commercially available DNA extraction platforms fo...

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Bibliographic Details
Published in:Journal of microbiological methods 2016-10, Vol.129, p.103-108
Main Authors: Burke, Rachael M., McKenna, James P., Cox, Ciara, Coyle, Peter V., Shields, Michael D., Fairley, Derek J.
Format: Article
Language:English
Subjects:
Achromopeptidase
Bacteria - isolation & purification
Bacteriological Techniques
Bacteriolysis
DNA, Bacterial - genetics
DNA, Bacterial - isolation & purification
Extraction
Feces - microbiology
Genome, Microbial
Glass
Group B Streptococcus
Humans
Microspheres
Polymerase Chain Reaction - methods
Reagent Kits, Diagnostic
Serine Endopeptidases - metabolism
Specimen Handling - methods
Streptococcal Infections - microbiology
Streptococcus
Streptococcus agalactiae - drug effects
Streptococcus agalactiae - genetics
Streptococcus agalactiae - isolation & purification
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Summary:Sub-optimal recovery of bacterial DNA from whole blood samples can limit the sensitivity of molecular assays to detect pathogenic bacteria. We compared 3 different pre-lysis protocols (none, mechanical pre-lysis and achromopeptidase pre-lysis) and 5 commercially available DNA extraction platforms for direct detection of Group B Streptococcus (GBS) in spiked whole blood samples, without enrichment culture. DNA was extracted using the QIAamp Blood Mini kit (Qiagen), UCP Pathogen Mini kit (Qiagen), QuickGene DNA Whole Blood kit S (Fuji), Speed Xtract Nucleic Acid Kit 200 (Qiagen) and MagNA Pure Compact Nucleic Acid Isolation Kit I (Roche Diagnostics Corp). Mechanical pre-lysis increased yields of bacterial genomic DNA by 51.3 fold (95% confidence interval; 31.6–85.1, p
ISSN:0167-7012
1872-8359
DOI:10.1016/j.mimet.2016.08.016